Heavy metal removal by a highly heavy metal tolerant sulfidogenic consortium in anaerobic semi-continuous stirred tank reactors (CSTR) [Elektronische Ressource] : changes of microbial community structure and abundance / Thi Quynh Hoa Kieu

Kieu

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• Publié par: technische_universitat_munchen
• Publié le: 01 janvier 2011
• Nombre de lectures: 31
• Poids de l'ouvrage: 1 Mo

Extrait

TECHNISCHE UNIVERSITÄT MÜNCHEN


Lehrstuhl für Siedlungswasserwirtschaft



Heavy metal removal by a highly heavy metal tolerant sulfidogenic
consortium in anaerobic semi-continuous stirred tank reactors (CSTR):
Changes of microbial community structure and abundance




Thi Quynh Hoa Kieu


Vollständiger Abdruck der von der Fakultät für Bauingenieur- und Vermessungswesen
der Technischen Universität München zur Erlangung des akademischen Grades eines
Doktors der Naturwissenschaften (Dr. rer. nat.)
genehmigten Dissertation.


Vorsitzender: Univ. - Prof. Dr.-Ing. Peter Rutschmann
Prüfer der Dissertation:
1. Univ. - Prof. Dr. rer. nat. Harald Horn
2. Univ. rer. nat. habil. Rudi F. Vogel




Die Dissertation wurde am 09.12.2010 bei der Technischen Universität München
eingereicht und durch die Fakultät für Bauingenieur- und Vermessungswesen am
20.01.2011 angenommen.
Thesis for the Degree of Doctor of Science




Heavy metal removal by a heavy metal tolerant sulfidogenic
consortium in anaerobic semi-continuous stirred tank reactors
(CSTR): Changes of microbial community structure

Advisor: Prof. Dr. rer. nat. Harald Horn













Kieu, Thi Quynh Hoa

Institute of Water Quality Control
Department of Civil Engineering and Geodesy
Technische Universität München
Germany, 2010 Table of Contents
Abstract..............................................................................................................................i
Acknowledgement............................................................................................................v
List of Tables ...................................................................................................................vi
List of Figures.................................................................................................................vii
List of Abbreviations ........................................................................................................x
Chapter 1. State of knowledge
1. Heavy metal pollution ..................................................................................................1
1.1. Heavy metal toxicity.........................................................................................1
1.2. Microbial resistance to toxic heavy metals.......................................................2
2. Heavy metal wastewater treatment methods ................................................................3
2.1. Chemical methods ............................................................................................3
2.2. Physico-chemical methods ...............................................................................3
2.3. Biological treatment methods...........................................................................4
3. Sulfate-reducing bacteria (SRB)...................................................................................5
3.1. Taxonomy and Phylogeny of sulfate-reducing bacteria ...................................7
3.2. Biochemistry of sulfate reduction.....................................................................8
3.3. Physiology of sulfate reduction ......................................................................10
3.3.1. Electron-donor metabolism ................................................................10
3.3.2. Electron-acceptor metabolism ............................................................10
3.3.3. Effect of pH to the activity of sulfate reducing bacteria.....................11
3.3.4. Effect of Eh to the activity 11
3.3.5. Effect of temperature to the activity of sulfate reducing bacteria ......11
4. Heavy metal removal by sulfate reduction ................................................................12
4.1. Passive treatment methods (or lime neutralization) .......................................12
4.2. Active treatment methods as sulfidogenic bioreactor.....................................13
5. Molecular biological approaches for analysis of microbial community wastewater
treatment .........................................................................................................................13
5.1. Microbial community analysis using rRNA as a molecular marker...............14 5.2. Full-cycle fingerprinting.................................................................................16
5.3. Nucleic acid fingerinting ................................................................................18
5.3.1. ARDRA (Amplified rDNA restriction analysis).................................19
5.3.2. DGGE/TGGE (Denaturing gradient gel electrophoresis/Temperature
Gradient gel electrophoresis)........................................................................19
5.3.3. Other fingerprinting techniques..........................................................21
5.4. FISH (Fluorescent in situ hybridization)........................................................23
6. Objectives and content of this study ..........................................................................25
7. References ..................................................................................................................26
Chapter 2. Heavy metal removal in batch conditions and in anaerobic semi-
continuous stirred tank reactors by a heavy metal tolerant sulfidogenic consortium
1. Introduction ................................................................................................................37
2. Materials and Methods ...............................................................................................39
2.1. Sulfidogenic consortia ....................................................................................39
2.2. Selection of a high heavy metal tolerance sulfidogenic consortium ..............39
2.2.1. Screening tests in test tubes................................................................40
2.2.2. Batch experiments ..............................................................................40
2.3. Continuous experiments .................................................................................41
2.3.1. Inoculum.............................................................................................41
2.3.2. Experimental set-up............................................................................41
2.3.3. Experimental procedure......................................................................43
2.3.4. Analytical methods .............................................................................43
3. Results ........................................................................................................................45
3.1. Screening tests in test tubes45
3.2. Batch experiments ..........................................................................................46
3.3. Semi-continuous experiments.........................................................................49
3.3.1. Heavy metal removal..........................................................................50
3.3.2. Effect of heavy metals on sulfate reduction and sulfide production ..51
3.3.3. Sulfur balance.....................................................................................51 3.3.4. Effect of heavy metals on sulfate-reducing bacteria population ........52
3.3.5. Effect of heavy metals on pH value ...................................................53
3.3.6. Qualitative EDS analysis....................................................................53
4. Discussion...................................................................................................................54
5. References57
Chapter 3. Application of different biomolecular techniques for assessing the
effect of heavy metals on microbial community structure in a heavy metal tolerant
sulfidogenic consortium
1. Introduction ................................................................................................................60
2. Materials and Methods ...............................................................................................61
2.1. Samples...........................................................................................................61
2.2. DNA extraction62
2.3. PCR amplication of 16S rRNA and dsrB genes .............................................62
2.4. Clone library analysis .....................................................................................63
2.5. DGGE of dsrB and 16S rRNA gene fragments ..............................................64
2.6. Phylogenetic analysis64
2.7. Totall cell counts and Fluorescent in situ hybridization (FISH).....................64
3. Results ........................................................................................................................65
3.1. Clone library analysis .....................................................................................65
3.2. DGGE of 16S rRNA and dsrB gene fragments ..............................................66
3.2.1. DGGE analysis of 16S rRNA gene fragments..........................